The quantity of amacrine cells and flapping frequency of pectoral fins in the transient ccdc141 and vstm2l knocked-out embryos. The statistical data showed that fewer th-expressing amacrine cells were observed in 4 dpf (A) ccdc141 and vstm2l exon 1 (E1) knocked-out embryos. A statistically significant hyperkinetic movements were found in 5 dpf (B) ccdc141 and vstm2l exon 1 knocked-out embryos. Fewer th-expressing amacrine cells were observed with statistical data in 4 dpf (C) ccdc141 and vstm2l exon 2 (E2) knocked-out embryos. Hyperkinetic movements were found statistically significant in 5 dpf (D) ccdc141 and vstm2l exon 2 knocked-out embryos, when compared to tracrRNA-Cas9 control. The not-injected AB wildtype embryos were used as no inj. control; for the exon 1-targeting knockouts in (A) and (B), 200 pg tracrRNA and 200 pg Cas9 protein were injected per embryo as basic tracrRNA-Cas9; basic tracrRNA-Cas9 and 50 pg ccdc141-crRNA E1/vstm2l-crRNA E1 were injected per embryo for ccdc141 exon 1/vstm2l exon 1 KO. For the exon 2-targeting knockouts in (C) and (D), 138 pg tracrRNA and 461 pg Cas9 protein were injected per embryo as basic tracrRNA-Cas9; basic tracrRNA-Cas9 and 34.6 pg ccdc141-crRNA E2/vstm2l-crRNA E2 were injected per embryo for ccdc141 exon 2/vstm2l exon 2 KO. Mann–Whitney U test was used for comparisons of unpaired nonparametric variables. All calculated P-values were two-tailed, and statistical significance was defined as P-value less than 0.05. Symbol meaning: *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001
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